Project description:Time course: Interaction of Puccinia hordei with Hordeum vulgare, Ingrid (leaf) and Puccinia triticana with Hordeum vulgare, Ingrid (leaf)
Project description:Small RNAs (21-24 nt) are pivotal regulators of gene expression that guide both transcriptional and post-transcriptional silencing mechanisms in diverse eukaryotes, including most if not all plants. MicroRNAs (miRNAs) and short interfering RNAs (siRNAs) are the two major types, both of which have a demonstrated and important role in plant development, stress responses and pathogen resistance. In this work, we used a deep sequencing approach (Sequencing-By-Synthesis, or SBS) to develop sequence resources of small RNAs from different Hordeum vulgare tissues (leaves, inflorescence and leaves inoculated with Blumeria). The high depth of the resulting datasets enabled us to examine in detail critical small RNA features as size distribution, tissue-specific regulation and sequence conservation between different organs in this species. We also developed database resources and a dedicated website (http://smallrna.udel.edu/) with computational tools for allowing other users to identify new miRNAs or siRNAs involved in specific regulatory pathways, verify the degree of conservation of these sequences in other plant species and map small RNAs on genes or larger regions of the maize genome under study.
Project description:Time course: Interaction of Magnaporthe isolate TH6772 (of the host plant rice) with Hordeum vulgare, Ingrid (leaf epidermis) and Magnaporthe isolate CD180 (of Pennisetum) with Hordeum vulgare, Ingrid (leaf epidermis)
Project description:Small RNAs (21-24 nt) are pivotal regulators of gene expression that guide both transcriptional and post-transcriptional silencing mechanisms in diverse eukaryotes, including most if not all plants. MicroRNAs (miRNAs) and short interfering RNAs (siRNAs) are the two major types, both of which have a demonstrated and important role in plant development, stress responses and pathogen resistance. In this work, we used a deep sequencing approach (Sequencing-By-Synthesis, or SBS) to develop sequence resources of small RNAs from different Hordeum vulgare tissues (leaves, inflorescence and leaves inoculated with Blumeria). The high depth of the resulting datasets enabled us to examine in detail critical small RNA features as size distribution, tissue-specific regulation and sequence conservation between different organs in this species. We also developed database resources and a dedicated website (http://smallrna.udel.edu/) with computational tools for allowing other users to identify new miRNAs or siRNAs involved in specific regulatory pathways, verify the degree of conservation of these sequences in other plant species and map small RNAs on genes or larger regions of the maize genome under study. Small RNA libraries were derived from leaves, inflorescence and leaves inoculated with Blumeria of Hordeum vulgare. Each tissue represented a mixture of developmental stages. Total RNA was isolated using the Plant RNA Purification Reagent (Invitrogen) and submitted to Illumina (Hayward, CA, http://www.illumina.com) for small RNA library construction using approaches described in (Lu et al., 2007) with minor modifications. The small RNA libraries were sequenced with the Sequencing-By-Synthesis (SBS) technology by Illumina. PERL scripts were designed to remove the adapter sequences and determine the abundance of each distinct small RNA. We thank Roger Wise for providing the plant material and Kan Nobuta and Gayathri Mahalingam for assistance with the computational methods..
Project description:Time course: Interaction of Blumeria graminis f. sp. hordei with Hordeum vulgare, Ingrid (leaf) and Blumeria graminis f. sp. tritici with Hordeum vulgare, Ingrid (leaf)
Project description:Time course: Interaction of Blumeria graminis f. sp. hordei with Hordeum vulgare, Ingrid (leaf epidermis) and Blumeria graminis f. sp. tritici with Hordeum vulgare, Ingrid (leaf epidermis)
Project description:RNA sequencing of the near-isogenic line mnd1.a in cv. Bowman background, compared to cv. Mesa and cv. Bowman Purpose: Expression analysis and variant calling.
